KR20190133724A - (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1 , 2-d] [1,4] oxazepin-9-yl) amino) propanamide, polymorphs and solid forms, and methods for preparing the same - Google Patents

Abstract

The present invention relates to (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] crystalline polymorphic form of imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide (GDC-0077), or stereoisomers, geometric isomers, tautomers thereof, or Pharmaceutically acceptable salts; And to a process for preparing said polymorphic form:

Description

(S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1 , 2-d] [1,4] oxazepin-9-yl) amino) propanamide, polymorphs and solid forms, and methods for preparing the same

Cross-reference of related applications

This application claims priority to US Provisional Application No. 62 / 491,812 (filed April 28, 2017), the contents of which are incorporated herein by reference in their entirety.

Technical Field

The invention provides (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] A polymorphic form of the PI3K inhibitor compound GDC-0077 referred to as polyzo [1,2-d] [1,4] oxazepin-9-yl) amino) propanamide. The invention also relates to a process for the preparation of the polymorphic form of GDC-0077.

Phosphoinositide 3-kinase (PI3K) is a lipid kinase that phosphorylates lipids at 3-hydroxy residues of the inositol ring (Whitman et al (1988) Nature, 332: 664). 3-phosphorylated phospholipids produced by PI3-kinase (PIP3) are kinases such as Akt and phosphinositide-dependent kinases by lipid binding domains (including the plekstrin homology (PH) region). Acts as a second messenger recruiting -1 (PDK1). The binding of Akt to the membrane PIP3 causes the translocation of Akt to the plasma membrane, resulting in contact of Akt with PDK1, which is responsible for activation of Akt. Tumor-inhibitor phosphatase (PTEN) dephosphorylates PIP3 to act as a negative regulator of Akt activation. PI3-kinase Akt and PDK1 are important in the regulation of many cellular processes, including cell cycle control, proliferation, survival, apoptosis and motility and are important elements of the molecular mechanisms of disease (eg cancer, diabetes and immune inflammation). Vivanco et al (2002) Nature Rev. Cancer 2: 489; Phillips et al (1998) Cancer 83:41).

The major PI3-kinase isoform in cancer is the class I PI3-kinase, p110 α (alpha) (US 5,824,492; US 5,846,824; US 6,274,327). Other isomorphisms are the cause of cardiovascular disease and immune inflammatory diseases (Workman P (2004) Biochem Soc Trans 32: 393-396); Patel et al (2004) Proceedings of the American Association of Cancer Research (Abstract LB-247 95th Annual Meeting, March 27-31, Orlando, Florida, USA; Ahmadi K and Waterfield MD (2004) Encyclopedia of Biological Chemistry (Lennarz WJ, Lane MD eds) Elsevier / Academic Press). The PI3 kinase / Akt / PTEN pathway is an attractive target for cancer drug development because these modulators or inhibitors are expected to inhibit proliferation, reverse the inhibition of apoptosis and overcome resistance to cytotoxic agents in cancer cells. (Folkes et al (2008) J. Med. Chem. 51: 5522-5532; Yaguchi et al (2006) Jour. Of the Nat. Cancer Inst. 98 (8): 545-556). PI3K-PTEN-AKT signaling pathway is deregulated in various cancers (Samuels Y, et al. (2004) Science 304 (5670): 554; Carpten J, et al (2007) Nature; 448: 439 -444]).

GDC-0077 (IUPAC name (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [ f] imidazo [1,2-d] [1,4] oxazepin-9-yl) amino) propanamide) also has strong PI3K activity (WO 2017/001645, US 2017/0015678, literature Edgar K. et al, # 156, "Preclinical characterization of GDC-0077, a specifıc PI3K alpha inhibitor in early clinical development" and Staben. S. # DDT02-0 "Discovery of GDC-0077, a highly isoform selective inhibitors of PI3Kalpha that promotes selective loss of mutant-p110alpha ", American Assoc. for Cancer Res. (AACR) annual meeting, April 2, 2017, Washington, DC]), and patients with locally advanced or metastatic solid tumors.

Many crystalline forms with different solid phase properties of drug substances can exhibit differences in bioavailability, shelf life and behavior during processing. Powder X-ray diffraction is an effective means of identifying different crystalline phases by unique diffraction patterns.

The pharmaceutical industry often faces the phenomenon of several polymorphs of the same crystalline chemical entity. Polymorphism often refers to the ability of a drug substance (ie, an active pharmaceutical ingredient (API)) to exist in two or more crystalline phases with different alignments and / or conformations of molecules in the crystal lattice to provide crystals with different physicochemical properties. It features. The ability to reliably produce selected polymorphic forms is an important factor in determining the success of drug products.

Global regulatory bodies require reasonable efforts to identify polymorphs of drug substances and to identify polymorphic interconversions. Due to the frequent unpredictable behavior of the polymorphs and the respective differences in their physicochemical properties, the consistency of manufacture between batches of the same product should be demonstrated. Proper understanding of the polymorphic landscape and the properties of the polymorphs of the drug will contribute to manufacturing consistency.

Crystal structure determination and intermolecular interactions at the atomic level provide important information for establishing absolute alignment (enantiomers), phase identification, quality control, and process development control and optimization. X-ray diffraction is widely recognized as a reliable means for crystal structure analysis and crystal form identification of pharmaceutical solids.

The utility of single crystals of drug substance is preferred because of the speed and accuracy of structure determination. However, it is not always possible to obtain crystals of suitable size for data collection. In this case, the crystal structure can be solved from the X-ray powder diffraction data obtained by measurement at ambient conditions and / or at various temperatures or humidity.

The present invention provides a polymorphic form of the PI3K inhibitor GDC-0077 of formula (CAS Registry No .: 2060571-02-8, Genentech, Inc.) ((S) -2-((2 -((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4 ] Oxazinpin-9-yl) amino) propanamide) or stereoisomers, geometric isomers, tautomers or pharmaceutically acceptable salts thereof:

An aspect of the invention is a pharmaceutical composition in the form of a polymorph of GDC-0077.

An aspect of the invention is a method of treating a hyperproliferative disorder of a mammal in the polymorphic form of GDC-0077.

An aspect of the present invention is a method for producing the crystalline polymorph of GDC-0077.

Embodiments of the invention provide (S) -2-((2-((S) -4- (die) designated as Form A polymorph, showing an X-ray powder diffraction pattern with characteristic peaks at about 5.7 ° 2-theta Fluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepin-9-yl) amino) It is a crystalline anhydride polymorph of propanamide. In some embodiments, the Form A polymorph exhibits an X-ray powder diffraction pattern having characteristic peaks at about 5.7, 11.4, and 19.0 ° 2-theta. In some embodiments, the Form A polymorph exhibits an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta.

Embodiments of the invention have a characteristic peak at about 5.7 ° 2-theta, or a characteristic peak at about 5.7, 11.4 and 19.0 ° 2-theta, obtained using an incident beam of Cu Kα radiation; (S) -2-((2-((S)-) designated as a Form A polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta 4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9- Crystalline anhydride polymorph of i) amino) propanamide.

Aspects of the invention are characteristic peaks at about 5.7 ° 2-theta, obtained using an incident beam of Cu Kα (1.541904 kV) radiation generated using Cross Beam optics (40 kV × 44 mA). Having; Has characteristic peaks at about 5.7, 11.4, and 19.0 ° 2-theta; (S) -2-((2-((S)-) designated as a Form A polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta 4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9- Crystalline anhydride polymorph of i) amino) propanamide.

An aspect of the invention is a Form A polymorph described herein that is substantially characterized by an X-ray powder diffraction pattern as shown in FIG. 4.

An aspect of the invention is the Form A polymorph described herein characterized by the X-ray powder diffraction peaks shown in Table 2.

An aspect of the invention is the Form A polymorph described herein wherein the differential scanning calorimetry (DSC) exhibits a melting endotherm at about 212 ° C to 215 ° C.

An aspect of the invention is the Form A polymorph described herein wherein the differential scanning calorimetry DSC exhibits melt endotherm at about 214 ° C.

Embodiments of the present invention are Form A polymorphs described herein that are substantially characterized by ¹³ C solid-state nuclear magnetic resonance (SSNMR) spectra as shown in FIG. 7A.

An aspect of the invention is a Form A polymorph described herein, substantially characterized by a ¹⁹ F SSNMR as shown in FIG. 7B.

Embodiments of the present invention specify (S) -2-((2-((S) designated as Form D polymorph, showing an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 10.8, 16.8 and 20.4 ° 2-theta. ) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine- Crystalline anhydride polymorph of 9-yl) amino) propanamide. In some embodiments, the Form D polymorph exhibits an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 8.6, 10.8, 16.8, 19.2, and 20.4 ° 2-theta. In some embodiments, Form D polymorphs described herein are characterized by the X-ray powder diffraction pattern shown in FIG. 15A. In some embodiments, Form D polymorphs described herein are characterized by the X-ray powder diffraction peaks shown in Table 3.

Embodiments of the invention have characteristic peaks at about 7.5, 10.8, 16.8 and 20.4 ° 2-theta, obtained using an incident beam of Cu Kα radiation; (S) -2-((2-((S)-) designated as a Form D polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 8.6, 10.8, 16.8, 19.2 and 20.4 ° 2-theta 4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9- Crystalline anhydride polymorph of i) amino) propanamide.

Aspects of the present invention are characteristic at about 7.5, 10.8, 16.8 and 20.4 ° 2-theta, obtained using incident beams of Cu Kα (1.541904 kPa) radiation generated using cross beam optics (40 kV × 44 mA). Has a peak; (S) -2-((2-((S)) designated as Form D polymorph, showing an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 8.6, 10.8, 16.8, 19.2, and 20.4 ° 2-theta -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9 -Yl) amino) propanamide is a crystalline anhydride polymorph.

Embodiments of the invention provide (S) -2-((2-((S) -4) designated as Form B polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 5.4, 10.5 and 25.2 ° 2-theta -(Difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepin-9-yl Crystalline trihydrate polymorph of) amino) propanamide. In some embodiments, the Form B polymorph exhibits an X-ray powder diffraction pattern with characteristic peaks at about 5.4, 10.5, 19.5, 20.1, 21.6, and 25.2 ° 2-theta. In some embodiments, Form B polymorphs described herein are characterized by the X-ray powder diffraction pattern shown in FIG. 12C. In some embodiments, Form B polymorphs described herein are characterized by the X-ray powder diffraction peaks shown in Table 2A.

Embodiments of the invention have characteristic peaks at about 5.4, 10.5 and 25.2 ° 2-theta, obtained using an incident beam of Cu Kα radiation; (S) -2-((2-((S)) designated as Form B polymorph, showing an X-ray powder diffraction pattern with characteristic peaks at about 5.4, 10.5, 19.5, 20.1, 21.6, and 25.2 ° 2-theta -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9 -Yl) amino) propanamide is a crystalline trihydrate polymorph.

Aspects of the invention feature characteristic peaks at about 5.4, 10.5 and 25.2 ° 2-theta, obtained using incident beams of Cu Kα (1.541904 kPa) radiation generated using cross beam optics (40 kV × 44 mA). Have; (S) -2-((2-((S)) designated as Form B polymorph, showing an X-ray powder diffraction pattern with characteristic peaks at about 5.4, 10.5, 19.5, 20.1, 21.6, and 25.2 ° 2-theta -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazepine-9 -Yl) amino) propanamide is a crystalline trihydrate polymorph.

Embodiments of the invention are pharmaceutical compositions comprising a therapeutically effective amount of the crystalline anhydride polymorph of Form A described above, and a pharmaceutically acceptable carrier, glidant, diluent or excipient.

An aspect of the present invention is a pharmaceutical composition comprising a therapeutically effective amount of the crystalline anhydride polymorph of Form D described above, and a pharmaceutically acceptable carrier, glidant, diluent or excipient.

An aspect of the present invention is a pharmaceutical composition comprising a therapeutically effective amount of the crystalline trihydrate polymorph of Form B described above, and a pharmaceutically acceptable carrier, lubricant, diluent or excipient.

An aspect of the invention is the pharmaceutical composition described above in the form of a tablet.

An aspect of the invention is the pharmaceutical composition described above, wherein the therapeutically effective amount is from about 1 to about 100 mg.

An aspect of the invention is the pharmaceutical composition described above, wherein the crystalline anhydride or trihydrate polymorph is milled.

Embodiments of the present invention provide (S) -2-((2-((S) -4- (difluoromethyl) -2) in ethanol (with or without water) or n-propanol (with or without water). Heating a slurry of oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide After the step, the mixture is cooled to have a characteristic peak at about 5.7 ° 2-theta; Has characteristic peaks at about 5.7, 11.4, and 19.0 ° 2-theta; Forming a Form A crystalline polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta. In some embodiments, the method comprises heating the slurry of GDC-0077 in ethanol in the presence of less than 40% (or less than 20% or less than 10%) of water to cool the mixture to form a Form A polymorph. Steps. In some embodiments, the method further comprises seeding the mixture with crystalline GDC-0077 (eg crystalline THF solvate).

Embodiments of the invention provide (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f ] Slurrying imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide in water (eg deionized water), (S) -2- ( (2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1 , 4] oxazinpin-9-yl) amino) propanamide. In some embodiments, the method comprises (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydro Slurrying Form A polymorph of benzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide in deionized water for 4 days at room temperature.

An aspect of the invention is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a crystalline polymorph of GDC-0077 (e.g., crystalline anhydride form A, crystalline anhydride form) D or crystalline trihydrate form B), or the crystalline polymorph of GDC-0077 described in detail herein (e.g., crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) and pharmaceutically acceptable carriers, glidants Administering a pharmaceutical composition comprising a diluent or excipient. In some embodiments, the cancer is an HR-positive and HER2-negative breast cancer expressing a PIK3CA mutation. In some embodiments, the method further comprises one or more additional therapeutic agents (eg, fulvestrant, palbociclib and / or letrozole).

Embodiments of the present invention provide a crystalline polymorph of GDC-0077 (eg, crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) described in detail herein for use in the treatment of cancer, or GDC described in detail herein. A pharmaceutical composition comprising -0077 crystalline polymorph (eg crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B). In some embodiments, the cancer is an HR-positive and HER2-negative breast cancer expressing a PIK3CA mutation. In some embodiments, the polymorph for use further comprises one or more additional therapeutic agents (eg, fulvestrant, palbociclib and / or letrozole). Embodiments of the present invention provide a crystalline polymorph of GDC-0077 (e.g., crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B), or GDC-0077, detailed herein, in the manufacture of a medicament for the treatment of cancer. Use of the pharmaceutical composition comprising a crystalline polymorph of (eg crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B). In some embodiments, the cancer is an HR-positive and HER2-negative breast cancer expressing a PIK3CA mutation. In some embodiments, the use comprises one or more additional therapeutic agents (eg, fulvestrant, palbociclib and / or letrozole).

Aspects of the invention are as described herein.

1 shows the XRPD pattern of the starting material GDC-0077.
2 shows the DSC and TGA traces of the starting material GDC-0077. Desolvation / evaporation endotherms and recrystallization endotherms occur at 64 ° C. and 141 ° C. (start), followed by melting endotherm at 214 ° C. TGA shows a weight loss of about 2.5% w / w before the melting event.
3 shows an overlay of the XRPD patterns of different solid form hits (forms I to VI) obtained in 96 well HTS polymorph screening for GDC-0077.
4 shows the XRPD of anhydride Form I (Form A) GDC-0077.
5A shows SEM images at 1000 × magnification of anhydride Form I (Form A) GDC-0077 (benchtop Phenom SEM (Nanoscience Instruments, Inc., USA)). Indicates.
FIG. 5B shows a PLM image of anhydride Form I (Form A) GDC-0077 (Leica DM 4000B microscope (Clemex Technologies Inc., Long, Quebec, Canada) with high resolution CCD camera and motorized stage. Gale material)), and 200X magnification).
6A shows thermal analysis of anhydride Form I (Form A) GDC-0077.
6B shows the TGA of Forms IV-VI.
6C shows the DSC of Forms IV-VI. In the DSC trace, multiple transitions, contributions to desolvation, formation of metastable forms and subsequent conversion to Form A and melting thereof were observed and indicated.
FIG. 6D shows the XRPD on the product obtained upon heating Form B (trihydrate) to 195 ° C., including the Form A pattern for comparison. As shown, the trihydrate ultimately converts to anhydride form A at this temperature.
FIG. 7A shows the ¹³ C SSNMR spectrum of Anhydride Form I (Form A) GDC-0077. FIG.
7B shows the ¹⁹ F SSNMR of Anhydride Form I (Form A) GDC-0077.
8 shows water sorption behavior of anhydride Form I (Form A) GDC-0077.
9 shows an isothermal TGA trace of trihydrate Form B GDC-0077 before and after equilibration at 60 ° C. at room temperature.
10A shows water sorption behavior of trihydrate Form B GDC-0077 at 25 ° C. FIG.
10B shows the DSC and TGA of Form III.
11 shows an overlay of XRPD patterns of Form A and Form III solid forms. Upon heating at room temperature and at 165 ° C. and 195 ° C., the XRPD pattern of Form III is shown. At higher temperatures (> 165 ° C.), Form III / C is converted to Form I / A.
FIG. 12A shows the conversion from anhydride (form A) to hydrate (form B) upon slurrying in deionized water for 4 days. Conversion starts within 12 hours as indicated by the hydrate marker peak (*) seen in the anhydride XRPD pattern. The mode change is completed by 96 hours. An XRPD pattern of Form B is included for reference.
12B shows slurry crosslinking experimental data of GDC-0077 hydrate-anhydride system in ethanol-water mixture at RT (room temperature). The equilibrium RH (relative humidity) band for the two forms was identified as 82-86%, which corresponds to 65-83% w / w water content.
12C shows the XRPD of trihydrate Form B GDC-0077.
13A shows the XRPD of GDC-0077 THF solvate.
13B shows thermal analysis of GDC-0077 THF solvate.
13C shows an overlay of the THF solvate (RT) and Form A XRPD pattern heated to 175 ° C. and 210 ° C. FIG. Solvates are desolvated to intermediate anhydride form, which ultimately converts to Form A.
FIG. 14 shows a thermal analysis of Form D (second anhydride form) obtained by desolvating THF solvate (polymorph of Form A). Phase transitions are indicated for each endotherm.
15A shows the XRPD of Anhydride Form D GDC-0077.
FIG. 15B is an overlay of the XRPD patterns of Forms A, D of GDC-0077 and final solid forms of GDC-0077 (obtained by slurrying Form A and D (1: 1 mixture) overnight in n-propanol (RT)) Indicates. Form D is converted to Form A in the slurry.
16A shows the PLM of milled GDC-0077. Form A is still stable when milling.
FIG. 16B shows DSC and MDSC tracking of GDC-0077 milled lots. Milling leads to the disorder demonstrated by the emergence of an exotherm (indicated by inset at 113 ° C.) indicating recrystallization of the disordered phase and subsequent melting of Form A (endotherm at 214 ° C.). MDSC does not show Tg close to exotherm.
17 shows the solid form landscape of GDC-0077.

Unless defined otherwise, technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs and are consistent with the following definitions:

Justice

The terms “comprises”, “comprises” are intended to specify the features, integers, components or steps mentioned when used herein and in the claims, but the presence of one or more other features, integers, components, steps or groups It does not make addition impossible.

As used herein, the term “about”, when used to refer to an X-ray powder diffraction pattern peak position, includes, for example, calibration of the instrument used, the process used to generate the polymorph, the crystallized material, and the like. Lifetime, refers to the inherent variability of peaks depending on the instrument used. In this case, the measurement variability of the instrument was about ± 0.2 ° 2-theta (θ). Those skilled in the art having the benefit of this disclosure may understand the use of "about" in the present application. The term “about” with respect to other defined parameters such as water content, C _max , t _max , AUC, intrinsic dissolution rate, temperature and time refers to the inherent variability in measuring or obtaining the parameters, for example. Those skilled in the art having the benefit of this disclosure can understand the variability of the parameters implied by the use of the term drug.

As used herein, "polymorph" refers to the presence of different crystalline forms of a compound that differ in packing or configuration / arrangement but have the same chemical composition. Crystalline forms have different alignments and / or conformations of molecules in the crystal lattice. Solvates are crystalline forms containing either stoichiometric or non stoichiometric amounts of solvent. If the solvent incorporated is water, the solvate is commonly known as a hydrate. Hydrates / solvates have the same solvent content, but lattice packing or coordination may exist as polymorphs for different compounds. Thus, a single compound may give rise to various polymorphic forms, where each form has different and distinct physical properties such as solubility profile, melting point temperature, hygroscopicity, particle shape, density, flowability, compatibility and / or X Has a line diffraction peak. The solubility of each polymorph can be different, and therefore the identification of the presence of the pharmaceutical polymorph is essential to providing a medicament with a predictable solubility profile. It is desirable to examine all solid forms of the drug, including all polymorphic forms, and determine the stability, dissolution and flow characteristics of each polymorphic form. Polymorphic forms of the compounds can be distinguished in the laboratory by X-ray diffraction analysis, or by other methods such as infrared, Raman or solid state NMR spectroscopy. For a general review of polymorphs and pharmaceutical applications of polymorphs, see G. M. Wall, Pharm Manuf. 3:33 (1986); J. K. Haleblian and W. McCrone, J. Pharm. Sci., 58: 911 (1969); ["Polymorphism in Pharmaceutical Solids, Second Edition (Drugs and the Pharmaceutical Sciences)", Harry G. Brittain, Ed. (2011) CRC Press (2009); And [J. K. Haleblian, J. Pharm. Sci., 64, 1269 (1975), all of which are incorporated herein by reference.

The acronym "XRPD" refers to X-ray powder diffraction, an analytical technique for measuring diffraction of X-rays in the presence of a solid component. Crystalline materials with regular repeating arrangements of atoms produce unique powder patterns. Materials with similar unit cells will provide a powder pattern with similar positions when measured in degrees 2θ (theta). Solvates exhibiting these properties are referred to as isostructures or isoform solvates. The intensity of the reflection varies with the electron density causing the diffraction, as well as the sample, sample preparation and instrument parameters. XRPD data analysis is based on the universal appearance of the powder pattern measured for known reactions of the X-ray diffraction system used for data collection. For diffraction peaks that may be present in the powder pattern, their position, shape, width and relative intensity distribution can be used to characterize the type of solid state order of the powder sample. The location, shape and intensity of any broad diffuse scattering (halo) on top of the mechanical background can be used to characterize the level and type of solid state order. The combined interpretation of the solid state order and the disorder provides a quantitative judgment of the macrostructure of the sample.

The term “package insert” is used to refer to instructions customarily included in the commercial package of a therapeutic product, which is used to indicate indications, instructions for use, dosage, administration, reasons for prohibition of use, and / or warnings about the use of such therapeutic products. Contains information about

As used herein, the term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable organic or inorganic salts of the compounds of the present invention. Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isicotinate, lactate, salicylate, acid sheet Laterate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharide, formate, Benzoate, glutamate, methanesulfonate (“mesylate”), ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (ie 1,1′-methylene-bis- (2-hydroxy-3 Naphthoate)) salts. Pharmaceutically acceptable salts may involve the inclusion of another molecule such as acetate ions, succinate ions or other counter ions. The counter ion can be any organic or inorganic moiety that stabilizes the charge of the parent compound. In addition, pharmaceutically acceptable salts may have more than one charged atom in its structure. If several charged atoms are part of a pharmaceutically acceptable salt, they may have several counter ions. For this reason, pharmaceutically acceptable salts may have one or more charged atoms and / or one or more counter ions.

If the compound of the present invention is a base, the preferred pharmaceutically acceptable salts can be prepared by any suitable method available in the art, for example by using the free base as an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methane. Sulfonic acids, phosphoric acid and the like, or organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid such as glucuronic acid or galacturonic acid, Alpha hydroxy acids such as citric or tartaric acid, amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as p-toluenesulfonic acid or ethanesulfonic acid and the like.

When the compound of the present invention is an acid, the preferred pharmaceutically acceptable salts can be prepared by any suitable method, for example by using a free acid as an inorganic or organic base, such as amines (primary, secondary or tertiary), alkali metal hydroxides or alkalis. It can be produced by treatment with earth metal hydroxide or the like. Illustrative examples of suitable salts include, but are not limited to, organic salts derived from amino acids (such as glycine and arginine), ammonia, primary, secondary or tertiary amines, or cyclic amines (such as piperidine, morpholine and piperazine), and sodium, Inorganic salts derived from calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

Preferred pharmaceutically acceptable salts can be prepared by any suitable method available in the art, for example by using the free base as an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid and the like, or organic acids. Such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid such as glucuronic acid or galacturonic acid, alpha hydroxy acid such as citric acid or tartaric acid , Amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as p-toluenesulfonic acid or ethanesulfonic acid and the like. Acids generally contemplated as being suitable for the formation of pharmaceutically useful or acceptable salts from basic pharmaceutical compounds are described, for example, in P. Stahl et al, Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; [S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66 (1) 1 19; [P. Gould, International J. of Pharmaceutics (1986) 33 201 217; Anderson et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York; Remington's Pharmaceutical Sciences, 18th ed., (1995) Mack Publishing Co., Easton PA; And The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference.

The term “pharmaceutically acceptable” indicates that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients included in the formulation and / or the mammal to be treated with it.

"Solvate" refers to an association or complex of one or more solvent molecules and a compound of the present invention. Examples of solvents forming solvates include, but are not limited to water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

The term "chiral" refers to a molecule having the non-superimposing properties of a mirror image partner, while the term "bichiral" refers to a molecule superimposed on a mirror image partner.

The term “stereoisomers” refers to compounds that have the same chemical composition but differ in the spatial arrangement of atoms or groups.

"Diastereoisomers" refer to stereoisomers having two or more chiral centers, the molecules of which are not mirror images of each other. Diastereomers have different physical properties such as melting point, boiling point, spectral properties and reactivity. Mixtures of diastereomers can be separated under high resolution analytical procedures (such as electrophoresis and chromatography).

"Enantiomer" refers to two stereoisomers of a compound that are mirror images that do not overlap each other.

Stereochemical definitions and conventions used herein are generally in accordance with the following: S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; And Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric or chiral centers and therefore may exist in different stereoisomeric forms. All stereoisomers of the compounds of the present invention, including but not limited to diastereomers, enantiomers, atropisomers and mixtures thereof, such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active form, that is, they have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L, or R and S, are used to indicate the absolute arrangement of the molecule relative to the chiral center of the molecule. The prefixes d and l, or (+) and (-), are used to indicate the rotational signal of plane-polarized light by the compound, where (-) or l means that the compound is left turnable. Compounds with the prefix (+) or d are right turnable. For the chemical structures shown, these stereoisomers are identical except that they are mirror images of one another. In addition, certain stereoisomers may be referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures. 50:50 mixtures of enantiomers are referred to as racemic mixtures or racemates, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species lacking optical activity.

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible through a low energy barrier. For example, proton tautomers (also known as protic tautomers) include interconversions through proton migration, such as keto-enol and imine-enamine isomerization. Valence tautomers include interconversions by reorganization of some of the bonding electrons.

Polymorph of GDC-0077

The invention provides (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] GDC of formula (I), referred to as multizo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide, which is incorporated herein by reference (WO 2017/001645, US 2017/0015678) Polymorph of -0077 (CAS reg. No .: 2060571-02-8), and processes, methods and reagents for the preparation of polymorph of GDC-0077:

As used herein, GDC-0077 includes all stereoisomers, geometric isomers, tautomers, and pharmaceutically acceptable salts thereof. GDC-0077 is an active pharmaceutical ingredient (API) in formulations being developed for the clinical treatment of breast cancer and other disorders.

X-ray powder diffraction analysis

Analysis of the X-ray powder diffraction (XRPD) pattern was performed using commercially available analytical software. XRPD is useful for fingerprinting different crystalline phases, polymorphs, hydrates or solvates by unique diffraction patterns. The so-called 2-theta values (the series of angles between the incident beam and the diffracted beam) are plotted along the abscissa (horizontal axis). The ordinate (vertical axis) records the intensity of the scattered X-rays recorded by the detector. The set of peaks acts as a unique fingerprint of the crystallographic unit cell in the crystalline material. A crystallographic unit cell is the smallest atomic-scale 3D fragment that repeats periodically in three dimensions over the entire crystal. All crystalline materials are distinguished by their crystallographic unit cells (and thus peak positions). By comparing the measured peak positions with those in the database, the crystalline material can be identified in a unique form. For pure materials, the positions of all peaks are generally defined by three parameters (a, b and c) and three angles (alpha, beta and gamma (α, β and γ)).

An XRPD pattern of the starting material GDC-0077 prepared as in Example 1 is shown in FIG. 1. As is evident from the increased baseline count and poorly resolved diffraction peaks, the starting material is poor crystalline. 2 shows the DSC (differential scanning calorimetry) and TGA (thermogravimetric analysis) traces of the starting material GDC-0077. Desolvation / evaporation endotherms and recrystallization endotherms occurred at 64 ° C. and 141 ° C. (starting), respectively, followed by melting endotherm at 214 ° C. TGA shows a weight loss of about 2.5% w / w before the melting event. TGA data (FIG. 2) show a weight loss of about 2.5% around 150 ° C. FIG. DSC thermograms show sharp melting endotherms with extrapolation onset temperatures of about 214 ° C., followed by significantly shallow endotherms (probably desolvation / evaporation) and exotherm (crystallization / realignment / phase conversion) in the range of 50 ° C. to 175 ° C. Indicates. Desolvation / evaporation endotherms and recrystallization endotherms occur at 64 ° C. and 141 ° C. (start), followed by melting endotherm at 214 ° C. TGA shows a weight loss of about 2.5% w / w before the melting event.

XRPD patterns of several different solid form hits were obtained from full plate (96 well) high speed mass screening as detailed in Example 2. 3 shows an overlay of the XRPD patterns of different solid form hits, Forms I-VI, obtained in 96 well HTS polymorph screening of GDC-0077. The two most frequently obtained forms were Forms I and II, of which Form II was consistent with the starting material (FIG. 3). Several other new polymorphic hits were identified from evaporation, precipitation and cooling plates, which were increased 10 × fold (ie 150-200 mg each for further characterization). Table 1 shows the growing conditions of six different Forms I-VI.

TABLE 1

Incremental conditions for high-speed mass screening of polymorph hits of GDC-0077

GDC-0077 solid form

4 shows the XRPD of anhydride Form I (Form A) GDC-0077. Table 2 shows the irradiation records of XRPD peaks of GDC-0077 Form I / A. 5A shows Scanning Electron Microscopy (SEM) at 1000 × magnification of Anhydride Form I (Form A) GDC-0077. 5B shows polarization microscopy (PLM) at 200 × magnification of anhydride Form I (Form A) GDC-0077. Form I was found to be substantially crystalline as small rod-shaped crystals of 30 to 40 μm in length. Thermal analysis of anhydride Form I (Form A) GDC-0077 shows negligible weight loss (about 0.25% w / w) by TGA and melting endotherm at 214 ° C. by DSC and is therefore considered anhydrous. (FIG. 6A). Compared to the experimental time scale, water sorption using the strict protocol mass change ratio (dm / dt) at 25 ° C. showed negligible water absorption of less than 0.3% w / w (Example 5). Since Form I is reasonably well characterized as being an anhydrous crystalline form, it is referred to as Form A.

TABLE 2

Record of XRPD peak survey of GDC-0077 Form I / A

Table 2 shows the peak irradiation record of the marker peak (with 31 peaks, maximum P / N = 152.2) of substantially crystalline Form I / A of GDC-0077 from the XRPD of FIG. 4: GMP lot @ pi = 136.1. Peak: 13 (pts) / Parabolic Filter, Threshold = 2.0, Cutoff = 5.0%, BG = 3 / 1.0, Peak-Top = Peak.

Forms IV through VI were shown to be similar due to differences in relative peak intensities in the 5-20 ° 2θ range by XRPD (FIG. 3). These three forms exhibited a weight loss of about 13% w / w up to 150 ° C. by TGA (FIG. 6B) and multiple transitions to melting point of 214 ° C. by DSC (FIG. 6C). These transitions are presumed to be after solvent loss (first endotherm, 88 ° C. for Forms IV and V), followed by crystallization of the intermediate form and subsequent melting / conversion to Form A (which melts at 214 ° C.). Form VI shows a similar tendency of recrystallization exotherm to Form A after desolvation by DSC. Multiple transitions, contributions to desolvation, formation of metastable forms and subsequent conversion to Form A and melting thereof were observed in the DSC traces. These transitions are shown in Figure 6c. The qNMR data for residual solvents determined that these forms were hydrates having a water content in the range of 13.4 to 13.8% w / w, which was in close agreement with the weight loss observed through the TGA. All three forms were heated to 195 ° C. in TGA and analyzed by XRPD and the product phase was identified as Form A (final stable anhydride form) (FIG. 6D). Since all three of these forms have been found to be stoichiometrically identical with very similar XRPD patterns that differ only in the relative intensity of the diffraction peaks, they are considered to be of the same form, ie trihydrate (3 moles of water / 1 mole of anhydride). Was considered, hereinafter referred to as Form B. XRPD obtained upon heating Form B (trihydrate) to 195 ° C. showed that the trihydrate ultimately converted to anhydride Form A at this temperature.

The desolvation behavior of Form B and its rehydration ability were further characterized. Thus, the compound was isothermally maintained at 60 ° C. in TGA until the weight loss was constant, followed by equilibration of the desolvated solid at 4 hours at room temperature, after which the experiment was repeated. 9 shows isothermal dehydration and weight loss profiles of equilibrated solids. In particular, FIG. 9 shows an isothermal TGA trace of trihydrate Form B GDC-0077 before and after equilibration at 60 ° C. at room temperature. As is evident from the data, the hydrate is easily dehydrated and loses almost all its lattice water (12%) at 60 ° C. and rehydrates after 4 hours of equilibrium, indicating easy passage of water into and out of the lattice. Water sorption desorption behavior of trihydrate Form B GDC-0077 at 25 ° C. is shown in FIG. 10A. Dynamic vapor sorption experiments provide information on the dehydration behavior of Form B. Dehydration of Form B begins rapidly below 40% RH (desorption curve) and completes by the time the sample is exposed to 0% RH. There is a hysteresis presenting an equilibrium lag, but anhydrous products are equally easily rehydrated above 40% RH. Unlike the resorption curves where the anhydride form shows a clear “surge” indicating anhydride-trihydrate conversion above 40% RH, desorption appears to be staged with a clear intermediate dehydrated form at 20-50% RH. Trihydrate dehydrates in two to three stages, whereas the formation of anhydride to hydrate occurs in one stage above 40% RH (25 ° C.).

The XRPD of Form III differs from Forms IV through VI (FIG. 3). Thermal analysis DSC and TGA traces of Form III of FIG. 10B were followed by recrystallization exotherm and final form (identified as Form A by XRPD) after several transitions (desolvation / evaporation endotherm) (similar to Form B of FIG. 6C). Melt is shown (FIG. 11). 11 shows an overlay of XRPD patterns of Form A and Form III solid forms. The XRPD pattern of Form III is shown at RT and upon heating to 165 ° C and 195 ° C. Form III converts to Form A upon heating above 165 ° C. (FIG. 11). Form III showed a weight loss of only 3% w / w by TGA, but its water content was found to be 11% by qNMR. On the other hand, Form II was found to be identical to the starting material characterized in FIGS. 1 and 2. Substantial similarity was observed in comparison of Form II, Form III and the XRPD pattern on the product obtained upon isothermal dehydration of Form B at 60 ° C., indicating that Forms II and III are simply partially dehydrated intermediates of Form B Only the degree of desolvation is different. The form is a partially dehydrated intermediate of Form B trihydrate. In addition, the possibility of the presence of the intermediate hydrate form is suggested from the staged desorption profile of Form B (FIG. 10A). For convenience of nomenclature, Form II / III (partially dehydrated form) is referred to as Form C. At higher temperatures (> 165 ° C.), Form III / C is converted to Form I / A.

From the characterization of the trihydrate (form B), it is clear that the trihydrate (form B) dehydrates to form A upon exposure to higher and / or lower relative humidity (RH), as shown in FIGS. 9, 10A and 6D. Do. The conversion from anhydride form A to form B was investigated. Dynamic vapor sorption (DVS) experiments do not exhibit thermodynamic conditions because these experiments are performed on a shorter time scale at which equilibrium cannot be obtained. To confirm hydrate formation, Form A was slurried in deionized water for 4 days (room temperature). As detected by XRPD using Form B as a reference, the mode conversion (from anhydride to hydrate) was initiated at 12 hours and completed within 4 days. The conversion from anhydride (form A) to hydrate (form B) was measured immediately after slurrying for 4 days in deionized water. Figure 12a began conversion within 12 hours as indicated by the hydrate marker peak at about 5.5 ° 2-theta shown in the anhydride XRPD pattern. The transition was completed about 96 hours.

12B shows the results of slurry crosslinking experiments of hydrate-anhydride mixtures, where water activity is plotted against vehicle composition (% water, v / v). 12B shows slurry crosslinking experimental data of GDC-0077 hydrate-anhydride system in ethanol-water mixture at RT (room temperature). Two forms of equilibrium RH (relative humidity) zones were identified as 82-86% corresponding to 65-83% w / w water content. The anhydride form (form A) was found to be stable at water activity (a _w ) of 0.82 or less, while the hydrate (form B) was found to be stable at a _w > 0.86. Thus, a _w of the anhydride-hydrate equilibrium ranges from 0.82 to 0.86.

These different water activity of the XRPD pattern of the solid obtained in the, anhydride / Form A is a hydrate / form a stable, while in a _w below 0.82 B exhibited stable at least a 0.86 _w.

12C shows the XRPD of trihydrate Form B GDC-0077. Table 2A shows the XRPD peak irradiation record of GDC-0077 Form B.

TABLE 2A

Record of XRPD peak survey of GDC-0077 trihydrate Form B

FIG. 13A shows the XRPD pattern of GDC-0077 THF solvate, which was found to contain 10% w / w THF when analyzed by LC-MS. A weight loss of 14% w / w was observed by TGA. DSC showed multiple transitions, which were further investigated by heating the samples to 175 ° C. and 201 ° C. before the endothermic generation (FIG. 13B). The overlay of the THF solvate and the XRPD pattern on the product obtained upon heating to 175 ° C. and 210 ° C. shows that the solvate dehydrates to the intermediate anhydride form, which ultimately converts to Form A (FIG. 13C). At 175 ° C. and as is clear from TGA, the THF solvate is fully dehydrated to form an intermediate anhydride form (Form D), which is then melted and recrystallized to Form A, which melts (starts) at 215 ° C. DSC and TGA traces of the anhydride form D (polymorph of Form A) in the second anhydride form obtained by dehydrating THF solvate are shown in FIG. 14. 14 shows the thermal analysis of Form D. Phase transitions are indicated for each endotherm. A negligible weight loss (less than 1% w / w) before melting confirms that Form D is anhydrous. As shown by DSC, this anhydride form melts at about 188 ° C. and after recrystallization to Form A, which melts at 215 ° C. Form D is also obtained by slurrying crude starting GDC-077 in n-propanol: water (99: 1 v / v) as the active pharmaceutical ingredient (API). THF solvates have been found to dehydrate to anhydride form D. Form D and Form A are involved in monotropic because Form A is a thermodynamically stable form over the range of temperatures studied. 15A shows the XRPD of Anhydride Form D GDC-0077. Table 3 shows representative XRPD peaks of Form D.

TABLE 3

XRPD peak survey recording of GDC-0077 Form D

Based on the "fusion heat rule" of Burger and Lamber's polymorphisms, two polymorphs are considered to be monotropic if those with higher melting points have greater heat of melting (Burger and Ramberger, Mikrochimica Acta (1979), 256]. In this case, depending on the sample history and purity, Form A melting at about 213 ° C. to 215 ° C. by thermal analysis of GDC-0077 THF solvate had a heat of fusion of about 100 J / g (FIG. 6A). Form D shows the onset of melting at 190 ° C. and a heat of fusion of about 82-48 J / g (FIG. 14). The ΔH (difference in fusion enthalpy or heat of fusion) of Form D cannot be accurately determined, but the value provides a suitable approximation to the rule, suggesting that the two forms are in monotropic. To further confirm this, slurry crosslinking experiments were performed in n-propanol at room temperature, which gave Form D from the raw API beforehand. FIG. 15B shows an overlay of the XRPD patterns of final solid forms of Form A, D and GDC-0077 (obtained after slurrying Form A and D (1: 1 mixture) overnight in n-propanol (room temperature)). Form D is converted to Form A in the slurry. 15B shows the XRPD pattern in solid form isolated from the slurry after 12 hours of stirring, which is consistent with that of Form A. This confirms that Form D is converted to Form A. In other words, Form A is a more stable form at room temperature to 214 ° C.

FIG. 17 shows the solid form landscape of GDC-0077 and provides a comprehensive snapshot of the different solid forms identified through high speed mass screening and during crystallization optimization. The shape landscape shown in the figures shows details of the phase transformations between the different shapes and the experimental conditions that enable these conversions as a guide to crystallization and augmentation of the appropriate shape. XRPD (FIG. 4) and solid state NMR (FIGS. 7A and 7B) confirmed Form A. FIG. 7A shows the ¹³ C SSNMR spectrum of Anhydride Form I (Form A) GDC-0077. FIG. 7B shows the ¹⁹ F SSNMR of Anhydride Form I (Form A) GDC-0077. Thermal analysis DSC and TGA traces are included in FIG. 6. Based on negligible weight loss and the absence of any dehydration event prior to melting at 212 ° C., Form A is identified as anhydride. Microscopy data (SEM and PLM) are shown in FIGS. 5A and 5B, where GDC-0077 form API particles appear to resemble plates. Table 4 shows particle size distribution (PSD) for 5, 15 and 30 second sonication. Water sorption of anhydride Form I (Form A) GDC-0077 is shown in FIG. 8. The compound absorbs negligible moisture (0.25% w / w) up to 90% RH (25 ° C).

TABLE 4

Particle size distribution (PSD) of GDC-0077 Form A as a function of sonication time

Effect of Particle Reduction on Form A

Milling of Form A may optimize certain PK properties. Enhanced Form A lots are milled and placed for stability studies (40 ° C./75% RH, 25 ° C./60% RH, open vials). An additional three lots were also milled, which behaved similarly during milling. Each milled lot was characterized and the physical shape was determined. This lot was slurried in 100% ethanol to obtain Form A and milled using a jet mill at 60 psi pressure for 3.5 hours. The yield was found to be 91%. Particle size analysis (PSD) determined the following: D10 = 0.7 μm, D50 = 2.7 μm, D90 = 6.7 μm. After milling, some API (active pharmaceutical ingredient, GDC-0077) batches were placed in stability at 40 ° C./75% RH and 25 ° C./60% RH in open vials. Solid phase data were collected at 4 and 8 weeks to assess the effect of temperature and humidity on physical form. Milled and stability samples were characterized by XRPD, PLM, DSC, TGA, moisture sorption analysis and surface area analysis. DSC runs were performed using an unsealed corrugated pan in a controlled mode, a heating rate of 1 ° C./minute from 0 ° C. to 175 ° C., a strain amplitude of ± 1 ° C. and a duration of 60 seconds.

To characterize the effect of milling through XRPD, thermal analysis and moisture sorption showed that the solid remained unchanged during milling, but the baseline count appeared to increase with decreasing peak resolution. The PLM image (FIG. 16A) shows that the crystallites are of uniform size and about 5 μm, confirming PSD data and demonstrating that milled GDC-0077 Form A remains stable during milling. Indications of disorder development are more evident in the DSC trace, where less endotherm is observed at 113 ° C. (inset in FIG. 16B) and melting endotherm of Form A is observed at 214 ° C. (FIG. 16B). This endotherm can contribute to the disorder, which is likely to be on the surface as in the case of milled material. Controlled Differential Scanning Calorimetry (MDSC) does not exhibit a glass transition temperature (Tg), but the absence of Tg does not rule out the possibility of surface amorphization, which suggests that highly mobile disordered surfaces tend to crystallize almost instantaneously at Tg. Because there is. The weight loss before melting is large in the milled material compared to the unmilled lot because of the disorder occurring as measured by thermogravimetric analysis (TGA). Milling produces a marked increase in surface area (10 ×) from 0.78 m ² / g to 6.68 m ² / g when milling (Table 5). The surface area determined by the BET analysis was found to be twice that produced simply taking into account the Sauter mean diameter (D3,2) hydrodynamic method of defining particle size, which was 3.42 m ² / g. This indicates that a marked increase in surface area is due to the occurrence of surface disorders. In addition, a 1% w / w increase in moisture uptake was observed in the water sorption profile of the milled lot (0.25% weight increase up to 90% RH, FIG. 8) compared to the unmilled material, which was disordered during milling. Further confirms the presence of.

TABLE 5

Surface Area of Milled GDC-0077 and Unmilled GDC-0077

The XRPD, DSC and TGA patterns of the milled samples at the time points T0, 4 and 8 show that the GDC-0077 physical form remains unchanged under stress-stable conditions (opened vials) as is apparent from XRPD. The reduction or loss of milling-induced disorder upon exposure to moisture is evident from the reduction of ΔH (25 ° C./60% RH) or complete absence of recrystallization endotherm (40 ° C./75% RH) in the stability samples by DSC. This is not unexpected because disordered phase annealing or recrystallization may occur upon exposure to moisture acting as a plasticizer. Thus, GDC-0077 Form A exhibits milling induced disorders, but the crystalline form remains unchanged upon size reduction. The disorder is believed to be surface related and reduced upon exposure to moisture and / or annealed in crystalline form. The physical form remains stable under open conditions for up to 8 weeks at 40 ° C./75% RH and 25 ° C./60% RH.

How to treat

The crystalline forms of GDC-0077 described in detail herein are useful for treating human or animal patients suffering from diseases or disorders (such as cancer) resulting from abnormal cell growth, function, or behavior associated with PI3K, and thus the GDC described in detail herein -0077 crystalline polymorph (eg crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) can be treated by a method comprising administering to the patient. In addition, a human or animal patient suffering from cancer can be treated by a method comprising administering to the patient a crystalline polymorph of GDC-0077 described in detail herein. This may improve or improve the patient's condition.

In addition, the method of the present invention is breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, skin cancer, keratinocyte tumor, lung cancer, dermoid carcinoma, large Cell carcinoma, non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenocarcinoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, undifferentiated carcinoma, papilloma carcinoma, normal carcinoma, melanoma, sarcoma, bladder Carcinoma, Liver Carcinoma & Bile Duct Cancer, Kidney Carcinoma, Pancreatic Cancer, Bone Marrow Disorders, Lymphoma, Hair Cell Cancer, Oral Cancer, Nasopharyngeal Cancer, Pharyngeal Cancer, Lip Cancer, Tongue Cancer, Oral Cancer, Small Intestine Cancer, Colon-Rectal Cancer, Colon Cancer, Rectal Cancer, Brain Cancer And central nervous system cancer, Hodgkin cancer, leukemia, bronchial cancer, thyroid cancer, liver cancer and hepatic bile duct cancer, hepatocellular carcinoma, gastric cancer, glioma / glioblastoma, endometrial cancer, melanoma, kidney cancer and renal cancer, bladder cancer, Uterine body cancer, cervical cancer, multiple myeloma And treating cancers selected from acute myeloid leukemia, chronic myeloid leukemia, lymphoid leukemia, chronic lymphocytic leukemia (CLL), myeloid leukemia, oral and pharyngeal cancer, non-Hodgkin's lymphoma, melanoma, and chorionic colon adenoma do.

Based on expression analysis, immunohistochemical analysis and cell line profiling, colon cancer, breast cancer, cervical cancer, gastric cancer, lung cancer and multiple myeloma respond best to PI3K modulators or inhibitors.

Combination therapy

Polymorphs of GDC-0077 may be used alone or in combination with additional therapeutic agents for the treatment of diseases or disorders described herein, such as inflammatory or hyperproliferative disorders (eg cancer). In certain embodiments, the crystalline polymorph of GDC-0077 (eg, crystalline anhydride form A, crystalline anhydride form D, or crystalline trihydrate form B) described in detail herein is an anti-inflammatory as a combination therapy in a pharmaceutical combination formulation or dosing regimen. Or in combination with additional second therapeutic compounds having anti-proliferative properties or useful for the treatment of inflammation, immune response disorders or hyperproliferative disorders (eg cancer). Additional treatments include CDK4 / 6 inhibitors, Bcl-2 inhibitors, JAK inhibitors, anti-inflammatory agents, immunomodulators, chemotherapy agents, apoptosis enhancers, nerve growth factors, cardiovascular disease drugs, liver disease drugs, antiviral drugs, blood disorder drugs, diabetes drugs And therapeutic agents for immunodeficiency disorders. The second therapeutic agent may be an NSAID anti-inflammatory agent. The second therapeutic agent may be a chemotherapy agent. The second compound of the pharmaceutical combination formulation or dosing regimen preferably has complementary activity against GDC-0077 so as not to adversely affect each other. Such compounds are suitably present in combination in amounts that are effective for the purpose intended. In one embodiment, a composition of the present invention is a crystalline polymorph of GDC-0077 described herein in detail in combination with a therapeutic agent such as a CDK4 / 6 inhibitor (eg crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B ).

Combination therapy may be administered as simultaneous or sequential curing. When administered sequentially, the combination may be administered in two or more administrations. Combination administration may include co-administration using separate formulations or a single pharmaceutical formulation, and continuous administration in any order, preferably with a period in which two (or all) active agents simultaneously exert their biological activity. Include.

Suitable dosages of any of the above co-administered formulations are those currently used and may be lowered due to the combined action (synergistic effect) of the newly identified formulation and other therapeutic agents or therapies.

Combination therapy can provide a "synergistic effect" and prove to be "synergistic", ie, the effect achieved when using the active ingredients together is greater than the sum of the effects resulting from the individual use of the compounds. The synergistic effect is that the active ingredient is: (1) co-formulated and administered or delivered in combined unit dosage form simultaneously; (2) delivered alternately or simultaneously as separate formulations; Or (3) when delivered by various other curing methods. When delivered in alternation therapy, a synergistic effect may be obtained when the compounds are administered or delivered sequentially, eg, by different injections of individual syringes, individual pills or capsules, or individual infusions. Generally, during alternation therapy, each active ingredient in an effective dosage is administered sequentially, ie continuously, while two or more active ingredients in an effective dosage in a combination therapy are administered together.

In certain embodiments of therapy, the crystalline polymorphs (eg, crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) of GDC-0077 described in detail herein may be incorporated into other therapeutic, hormonal or antibody preparations (such as And those combined with surgical and radiation therapy as well. Thus, combination therapies according to the present invention may be used for the administration of crystalline polymorphs (eg, crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) of GDC-0077 described in detail herein, and one or more other methods of treating cancer. Includes the use of. The amount of crystalline polymorph and pharmaceutically active therapeutic agent of GDC-0077 and the relative timing of administration can be selected to achieve the desired combined therapeutic effect.

Additional therapeutic agents used in combination with crystalline polymorphs (eg, crystalline anhydride Form D, crystalline anhydride Form D, or crystalline trihydrate Form B) of GDC-0077 described in detail herein are 5-FU, docetaxel, erybulin, gemcitabine , Cobimetinib, ipatasetip, paclitaxel, tamoxifen, fulvestrant, GDC-0810, dexamethasone, palbociclib, bevacizumab, pertuzumab, trastuzumab emtansine, tsastuzumab and letrozole Include.

In some embodiments, an individual in need thereof is administered a therapeutically effective amount of a crystalline polymorph of GDC-0077 (eg, crystalline anhydride form A, crystalline anhydride form D, or crystalline trihydrate form B) described in detail herein. Provided is a method of treating cancer in a subject in need thereof. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is hormone receptor positive (HR +) breast cancer. In some embodiments, the cancer is estrogen receptor positive (ER +) breast cancer. In some embodiments, the cancer is HER2-negative breast cancer. In some embodiments, the cancer is HR + metastatic breast cancer. In some embodiments, the cancer is HR-positive, HER2-negative advanced breast cancer. In some embodiments, the cancer is HER2-negative, ER-negative and progesterone receptor (PR) -negative breast cancer. In some embodiments, the individual is a human. In some embodiments, the individual is a postmenopausal woman. In some embodiments, the breast cancer subtype is Basal or Luminal. In some embodiments, the cancer has a PIK3CA mutation. In some embodiments, the cancer expresses a PIK3CA mutant selected from E542K, E545K, Q546R, H1047L, and H1047R. In some embodiments, the cancer expresses a PTEN mutant.

In some of these embodiments, the method of treating cancer (eg, breast cancer) further comprises administering to the individual one or more additional therapeutic agents. In some embodiments, one or more additional therapeutic agents are CDK4 / 6 inhibitors (eg, palbociclib, ribosiclip, and abemashic), selective estrogen receptor inhibitors (SERD) (eg fulvestrant), and aromas Other agent inhibitors (eg letrozole). In some embodiments, the additional therapeutic agent is palbociclib. In some embodiments, the additional therapeutic agent is fulvestrant. In some embodiments, one or more additional therapeutic agents are palbociclib and letrozole.

Embodiments of the present invention provide a crystalline polymorph of GDC-0077 (eg, crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) described in detail herein for use in the treatment of cancer, or GDC described in detail herein. A pharmaceutical composition comprising -0077 crystalline polymorph (eg crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B). In some embodiments, the cancer is an HR-positive and HER2-negative breast cancer expressing a PIK3CA mutation. In some embodiments, the polymorph for use further comprises one or more additional therapeutic agents (eg, fulvestrant, palbociclib and / or letrozole).

Embodiments of the present invention provide a crystalline polymorph of GDC-0077 (e.g., crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B) as described herein in detail in the manufacture of a medicament for the treatment of cancer, or as described herein in detail. Use of a pharmaceutical composition comprising a crystalline polymorph of GDC-0077 described (eg, crystalline anhydride form A, crystalline anhydride form D or crystalline trihydrate form B). In some embodiments, the cancer is an HR-positive and HER2-negative breast cancer expressing a PIK3CA mutation. In some embodiments, the use further comprises one or more additional therapeutic agents (eg fulvestrant, palbociclib and / or letrozole).

Pharmaceutical Compositions and Formulations

The polymorphic form of GDC-0077 of Formula I is formulated according to standard pharmaceutical practice for use in therapeutic combinations for therapeutic treatment (including prophylactic treatment) of hyperproliferative disorders of mammals (including humans). Can be converted. The present invention provides a pharmaceutical composition comprising GDC-0077 jointly with one or more pharmaceutically acceptable carriers, lubricants, diluents or excipients.

Suitable carriers, diluents, glidants and excipients are well known to those skilled in the art and include materials such as carbohydrates, waxes, water soluble and / or swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like.

The formulations may be prepared using conventional dissolution or mixing procedures. The compounds of the present invention are typically formulated in pharmaceutical dosage forms to provide an easily controllable dosage of the drug and to allow the patient to comply with the prescribed regimen.

The pharmaceutical composition (or formulation) for the application may be packaged in various ways depending on the method used for drug administration. Generally, the dispensing article includes a container having the pharmaceutical formulation deposited therein in a suitable form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, metal cylinders and the like. In addition, the container may include an easily manipulated ensemblar that prevents indiscriminate access to the contents of the package. It also has a label deposited on top that describes the contents of the container. In addition, the label may include appropriate warnings.

Pharmaceutical formulations in the form of polymorphs of GDC-0077 are formulated for various routes and types of administration together with pharmaceutically acceptable diluents, carriers, excipients, glidants or stabilizers in the form of lyophilized formulations, milled powders or aqueous solutions. (Remington's Pharmaceutical Sciences (1995) 18th edition, Mack Publ. Co., Easton, PA). Formulation may be carried out by mixing with a physiologically acceptable carrier (ie, a carrier that is nontoxic to the recipient at the dosages and concentrations employed) at the desired degree of purity at appropriate pH at ambient temperature. The pH of the formulation mainly depends on the particular use and concentration of the compound, but may range from about 3 to about 8.

The pharmaceutical formulation is preferably sterile. In particular, formulations to be used for in vivo administration must be sterile. Such sterilization can be readily accomplished by filtration through sterile filtration membranes.

Pharmaceutical formulations can usually be stored as solid compositions, tablets, pills, capsules, lyophilized formulations, or as aqueous solutions.

The pharmaceutical formulation of the present invention will be administered and administered according to the amount, concentration, schedule, procedure, vehicle and route of administration consistent with good medical practice. Factors considered herein include the particular disorder to be treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the formulation, the method of administration, the schedule of administration and other factors known to the physician.

Acceptable diluents, carriers, excipients and stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate and other organic acids; Antioxidants such as ascorbic acid and methionine; Preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzetonium chloride; phenol, butyl, ethanol, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol Cyclohexanol; 3-pentanol; and m-cresol); Low molecular weight (less than about 10 residues) polypeptides; Proteins such as serum albumin, gelatin or immunoglobulins; Hydrophilic polymers such as polyvinylpyrrolidine; Amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; Monosaccharides, disaccharides and other carbohydrates such as glucose, mannose or dextrins; Chelating agents such as EDTA; Sugars such as lactose, sucrose, mannitol, trehalose or sorbitol; Salt-forming counter ions such as sodium; Metal complexes (eg Zn-protein complexes); And / or nonionic surfactants such as TWEEN® such as Tween 80, PLURONICS® or polyethylene glycol (PEG) such as PEG400. In addition, the active pharmaceutical ingredient may be prepared, for example, by microcapsules prepared by coacervation or interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly- (methylmethacrylate) microcapsules respectively). , Colloidal drug delivery systems (eg liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or microemulsions. Such techniques are described in Remington's Pharmaceutical Sciences 18th edition, (1995) Mack Publ. Co., Easton, PA. Other examples of drug formulation can be found in Lierman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, Vol 3, 2nd Ed., New York, NY.

Tablets may include one or more pharmaceutically acceptable carriers, glidants, diluents or excipients selected from microcrystalline cellulose, lactose, sodium starch glycolate and magnesium stearate.

Pharmaceutically acceptable glidants include silicon dioxide, powdered cellulose, microcrystalline cellulose, metallic stearate, sodium aluminosilicate, sodium benzoate, calcium carbonate, calcium silicate, corn starch, magnesium carbonate, asbestos talc, stearose It can be selected from wet C, starch, starch 1500, magnesium lauryl sulfate, magnesium oxide and mixtures thereof.

Pharmaceutical formulations include those suitable for the route of administration described in detail herein. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Techniques and formulations are generally described in Remington's Pharmaceutical Sciences 18th Ed. (1995) Mack Publishing Co., Easton, PA. The method includes the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. Generally, formulations are prepared by associating the active ingredient with a liquid carrier or finely divided solid carrier or both uniformly and densely, followed by shaping the product as necessary.

The pharmaceutical compositions may be in the form of sterile injectable preparations, such as sterile injectable aqueous or oily suspensions. Such suspensions may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. Sterile injectable preparations may be solutions or suspensions in nontoxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol or lyophilized powder. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils may conveniently be used as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables.

Administration of the pharmaceutical composition

The pharmaceutical composition of the present invention may be administered by any route suitable for the disease to be treated. Suitable routes include oral, parenteral (including subcutaneous, intramuscular, intravenous, intraarterial, inhalation, intradermal, intradural, epidural, and infusion techniques), transdermal, rectal, nasal, topical (ball and sublingual) Vaginal, intraperitoneal, pulmonary and intranasal routes. In addition, topical administration may involve transdermal administration, such as the use of transdermal patches or iontophoretic devices. Formulation of the drug is discussed in Remington's Pharmaceutical Sciences, 18th Ed., (1995) Mack Publishing Co., Easton, PA. Other examples of drug formulation can be found in Lierman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, Vol 3, 2nd Ed., New York, NY. For topical immunosuppressive treatment, the compounds can be administered by intralesional administration (including spraying) or by contacting the graft with the inhibitor prior to transplantation. It will be appreciated that the preferred route may vary with for example the condition of the recipient. When the compound is administered orally, it may be formulated into pills, capsules, tablets and the like together with pharmaceutically acceptable carriers, glidants or excipients. When the compound is administered parenterally, it may be formulated in unit dosage form with a pharmaceutically acceptable parenteral vehicle or diluent as described in detail below.

Dosages for treating human patients range from polymorphic forms of GDC-0077, such as from about 2 to about 50 mg, about 3 to about 20 mg, about 3 to about 15 mg, about 3 to about 100 mg. About 20 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 9 mg, about 12 mg, about 15 mg, or about 20 mg of the compound. Dosage is once daily (QD), twice daily (BID), or more frequently, depending on pharmacokinetic (PK) and pharmacodynamic (PD) properties (including absorption, distribution, metabolism and excretion of specific compounds). can do. In addition, virulence factors can affect dosage and dosing regimen. When administered orally, pills, capsules or tablets may be taken twice daily, daily or less frequently, such as once weekly, or once every two or three weeks, for a specified period of time. Curing may be repeated for many cycles of therapy.

Example

Example 1: (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] Isolation and Physicochemical Characterization of Imidazo [1,2-d] [1,4] oxazepin-9-yl) amino) propanamide (GDC-0077)

GDC-0077 was prepared according to WO 2017/001645, US 2017/0015678, each of which is incorporated by reference.

(S) -3- (9-bromo-5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazin-2-yl) -4- (difluoro Methyl) oxazolidin-2-one (600 mg, 1.50 mmol), L-alanine (267 mg, 3.00 mmol), cuprous iodide (57 mg, 0.30 mmol) and tertiary potassium phosphate (637 mg, 3.00 mmol) Was suspended in dimethyl sulfoxide (6.0 mL). The reaction mixture was heated at 100 ° C. for 2 hours. Cool to rt and add dimethyl sulfoxide (4.0 mL), ammonium chloride (480 mg, 9.00 mmol) and triethylamine (3.1 mL, 22.5 mmol). To the resulting stirred suspension 1- [bis (dimethylamino) methylene] -1H-1,2,3-triazolo [4,5-b] pyridinium 3-oxide hexafluorophosphate (5.10 g, 13.5 mmol) was added in portions for 5 minutes. The reaction mixture was stirred at rt for 1 h, then filtered through Celite® and washed with ethyl acetate. The organic extract was washed with saturated aqueous sodium bicarbonate and the aqueous phase was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo. The crude residue was purified via silica gel flash chromatography (solvent gradient: 0-5% methanol in dichloromethane) followed by chiral supercritical fluid chromatography to give GDC-0077 (294 mg, 46%) as an off-white solid. It was. LCMS (ESI): R _T (min) = 2.89 [M + H] ⁺ = 408, Method = A; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.00 (d, J = 8.7 Hz, 1H), 7.38 (br s, 1H), 7.18 (s, 1H), 7.00 (br s, 1H), 6.71 ( t, J = 55.9 Hz, 1H), 6.41 (dd, J = 8.8, 2.3 Hz, 1H), 6.16 (d, J = 7.2 Hz, 1H), 6.09 (d, J = 1.9 Hz, 1H), 5.02- 4.89 (m, 1H), 4.63-4.52 (m, 2H), 4.39-4.30 (m, 4H), 3.76 (quintet, J = 7.0 Hz, 1H), 1.30 (d, J = 7.1 Hz, 3H).

GDC-0077 Form A (anhydride) was obtained by slurrying GDC-0077 in ethanol at 50 ° C. for 4 hours, then evaporating the solvent under nitrogen purge to give highly crystalline Form A. DSC thermograms showed one endothermic transition starting at about 212 ° C. to 214 ° C. and associated heat of fusion of about 107 J / g. The water solubility of Form A at room temperature was 30.8 μg / mL at pH 7.06. Form A was slurried in deionized water for 4 days (room temperature), the slurry was centrifuged to remove the supernatant, and then the solid was dried at room temperature for several hours to obtain trihydrate form.

After dry granulating Form A GDC-0077 using a roller compact, a purification operation was performed. Additional ingredients of the tablets include microcrystalline cellulose (Avicel® PH 102, FMC BioPolymer), lactose (FastFlo® 316, Foremost Farms USA). ), Sodium starch glycolate (EXPLOTAB®, JRS Pharma) and magnesium stearate (Hyqual®, Macro Fine Chemicals) Included.

Example 2: High Speed Mass Polymorph Screening (HTS)

Automated HTS (Freeslate Inc., Calif.) Of 96-well plates using the Symyx CM2 system was performed to identify potential polymorphic forms of GDC-0077. About 20 mg of API was added to each well, to which 800 μl of solvent (pure or mixture) was added and the slurry was stirred at 50 ° C. for 2 hours. The solvent is water, 1,2-dichloroethane, heptane, cyclohexane, ethanol, 1-propanol, acetonitrile, butylamine, nitromethane, 1,4-dioxane, benzene, perfluoroheptane, ethyl acetate, (tri Fluoromethyl) benzene, butan-2-one (MEK), 1,2-dimethoxyethane, 2-methyltetrahydrofuran, carbon tetrachloride, dimethylacetamide, tetrahydrofuran (THF), acetone, anisole , Toluene and 2-ethoxyethanol. From this "master" plate, the supernatant was filtered and partitioned into three separate plates for evaporation, precipitated by anti-solvent addition and controlled cooling was performed at 50 ° C.-20 ° C. for 8-10 hours. Details on solvents and anti-solvents are shown in FIG. 2. In all cases, residual solvents were evaporated or removed by siphon and solids were tested using polarization microscopy and X-ray powder diffraction analysis. After comparing the XRPD patterns of crystalline hits, there was potentially an increase in "different" hits and their characterization.

Example 3: Slurry Crosslinking

Deionized water (DI) -ethanol (anhydrous alcohol) mixtures of different compositions of 0 to 100% water were prepared and their water activity was measured using a water activity meter. Slurry experiments were set up at room temperature where a 1: 1 mixture (40 mg total) in the form of trihydrate and anhydride of GDC-0077 was added to the solvent mixture (1.5 mL liquid) and stirred at room temperature for 4 days. After 4 days, samples were aliquoted and centrifuged. Solids were analyzed by XRPD and supernatants were analyzed for water activity. A 1: 1 mixture of forms A and D was slurried in n-propanol overnight at room temperature. The slurry was centrifuged and the solid form was analyzed by XRPD.

Example 4: Ambient X-Ray Powder Diffraction Analysis (XRPD)

An XRPD pattern was prepared using a Rigaku SmartLab® diffractometer (Rizaku Corporation) using an incident beam of Cu Kα (1.541904 μs) radiation generated using cross beam optics (40 kV × 44 mA). Rigaku Corp., Tokyo, Japan). GDC-0077 powder samples are packed on a zero-background holder using the top fill method and the scan is 2-40 ° in a Bragg-brentano or parallel beam array (reflection geometry). Acquired at a scan rate of 1 or 3.0 ° / min and a step size of 0.02 or 0.04 ° 2θ (2-theta) over the 2θ range. Data was analyzed using commercial software (JADE®, Version 9, Materials Data Inc., Livermore, CA, USA).

Example 5: Moisture Sorption Analysis

About 5-6 mg of powder sample was placed in a sample pan of an automated moisture sorption analyzer (Q5000SA, TA instruments, New Castle, Delaware, USA) at 25 ° C. and a nitrogen flow rate of 200 mL / min. The sample was initially "dried" at 600% 0% RH for a total of 60 minutes (60 to 25 ° C), then gradually increasing RH from 0% to 90% in 10% increments with a 240 minute hold time The dm / dt window was 0.001% for 30 minutes at each RH. The RH was then gradually reduced back to 0% RH with a reduction of 10% using the same protocol. For the hydrate samples, the treatment was reversed, where the starting RH was maintained at 90%, then gradually decreased to 0% and then similarly increased back to 90%. This was done so that the water of hydration was maintained at the beginning of the experiment.

Example 6: Moisture Activity

Aqualab 4TEV (Decagon Devices, Washington, USA) was used as a water activity meter in capacitive sensor mode to obtain data for solvent mixtures and slurry supernatants at 25 ± 0.2 ° C. . The instrument was calibrated using vendor supplied reference (saturated salt solution) over the a _w range of 0.25-1. All a _w values were obtained after stabilization of three consecutive reads.

Example 7: Differential Scanning Calorimetry (DSC)

Approximately 3-8 mg of powder samples were analyzed using DSC Q2000TM (TA Instruments, New Castle, Delaware, USA) with refrigerated refrigeration accessories. Samples were packed in unsealed pans (Tzero, aluminum pans) and typically heated from 20 ° C. to 250 ° C. under a dry nitrogen purge. The instrument was calibrated using sapphire (base) and indium (temperature and cell constant). The data was analyzed using commercial software (Universal Analysis 2000, Version 4.7A, TA Instruments). Experimental conditions and fan arrangement are as follows:

Example 8: Thermogravimetric Analysis (TGA)

Non-isothermal experiments: In a thermogravimetric analyzer (Discovery TGA, T. Instruments), 3-4 mg of GDC-0077 sample was heated at 350 ° C. from room temperature to 10 ° C./min in an open aluminum pan. Heated and heated to 350 ° C. at room temperature under a nitrogen purge. Temperature calibration was performed using Alumel® mill nickel. Standard weights of 100 mg and 1 g were used for the weight calibration.

Isothermal Experiment: In a thermogravimetric analyzer (Q500 TGA, TAI Instruments), 3-4 mg of GDC-0077 samples were heated in an open aluminum pan at room temperature at 60 ° C. at a heating rate of 10 ° C./min and isothermal at 60 ° C. overnight. Was maintained. The sample was then analyzed by XRPD or left at equilibrium in the sample pan for 4 hours at room temperature, then repeated isothermally at 60 ° C. using the same experimental parameters as mentioned above.

Example 9: Polarization Microscopy (PLM)

Samples were dispersed in silicone oil and observed under a cross polarizer of a video enhanced Leica DM 4000B microscope (Clemex Technologies, Inc., Longale, Quebec, Canada) with a high resolution CCD camera and a motorized stage at 200 × magnification. Photomicrographs were acquired using Claymex Vision PE software (Clemex Technologies, Inc., Long Gale, Quebec, Canada).

Example 10 Scanning Electron Microscopy (SEM)

After the powder sample sputter was coated onto the SEM stub, it was tested using a benchtop Phenom SEM (NanoSciences Instruments, Arizona, USA). Micrographs were obtained at different magnifications.

Example 11: Particle Size Distribution Analysis (PSD)

Particle size analysis was performed using a Malvern Mastersizer 2000 instrument (Malvern Instruments Ltd., Malvern, UK) with a Hydros 2000SM wet dispersion attachment. About 30 mg of API was weighed into a vial and 0.1% Span 85 in 1 mL of heptane was added. The vial was sonicated for 5 seconds, about 0.3 mL was added to the sampler at a stirring speed of 1500 rpm, and the PSD was performed at 10-20% concealment. The sample was then sonicated for an additional 10 seconds (15 seconds total), about 0.3 mL was added to the sampler, a PLM image was obtained, and the PSD was performed. The same sample was then sonicated for an additional 15 seconds (30 seconds total), about 0.3 mL was added, a PLM picture was taken, and a PSD was obtained. From the sonication study, the appropriate sonication period was selected. PLM images and PSDs were used to determine the amount of sonication required for the sample to disperse the flock but prevent or minimize crystal breakage. About 10 mg additional three samples were weighed into vials and 0.1% span 85 in 1 mL heptane was added. Samples were sonicated during the sonication period determined in the sonication study. Final PSD analysis was performed in three experiments using a predetermined sonication period. The instrument was treated twice with isopropyl alcohol (IPA) and once heptane, then charged with 0.1% span 85 in heptane for each sample. After running the last sample, the instrument was rinsed once with IPA.

Example 12 Surface Area Analysis

Surface area measurements were performed using a Micromeritics ASAP 2460 (Micromeritics Instrument Corp., Georgia, USA) equipped with a Micromeritics Smart VacPrep attachment. 500 mg to 1 g of sample was weighed into an empty ASAP 2460 tube, placed in a smart backprep and degassed under ambient conditions for 24 hours, and then exposed to krypton gas adsorption at 25 ° C. and 100 mm Hg holding pressure. Eleven-point measurements were performed at relative pressures ranging from 0.050 to 0.300 and data were analyzed using MicroActive software provided by the supplier.

Example 13: Solid-State Nuclear Magnetic Resonance Spectroscopy (SSNMR)

All ¹³ C (@ 8 kHz spinning rate) SSNMR experiments were performed using a 500 MHz Bruker instrument (Bruker BioSpin GmbH, Karlsruhe, Germany). ¹³ C data was obtained using CP / TOSS sequence. 1-2 K scans were collected for signal averaging. A contact time of 4 ms and a recycle delay of 5 seconds were used. Spinal 64 sequences were used for decoupling with a pulse length of 5.3 μs. A ¹ H 90-degree pulse length of 2.9 μs was used. All ¹⁹ F (@ 14 kHz spinning rate) SSNMR experiments were performed using a 500 MHz Bruker instrument. ¹⁹ F data were obtained using CP sequences. 64 to 256 K scans were collected for signal averaging. A contact time of 750 μs and a recycle delay of 7 seconds were used. A ¹ H 90-degree pulse length of 3.54 μs was used.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the description and examples should not be construed as limiting the scope of the invention. Accordingly, all suitable modifications and equivalents may be considered to be within the scope of the invention as defined by the following claims. The disclosures of all patent and scientific literature cited herein are expressly incorporated by reference in their entirety.

Claims (26)

(S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidine-) showing an X-ray powder diffraction pattern with characteristic peaks at about 5.7 ° 2-theta Form A polymorph which is a crystalline anhydride polymorph of 3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide. The method of claim 1,
Form A polymorph showing an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, and 19.0 ° 2-theta. The method according to claim 1 or 2,
Form A polymorph showing an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta. The method according to any one of claims 1 to 3,
Form A polymorph characterized by the X-ray powder diffraction pattern shown in FIG. 4. The method according to any one of claims 1 to 3,
Form A polymorph characterized by an X-ray powder diffraction peak shown in Table 2. The method according to any one of claims 1 to 5,
Form A polymorph with differential scanning calorimetry (DSC) showing melt endotherm at 212 ° C to 215 ° C. The method according to any one of claims 1 to 6,
Form A polymorph, characterized by ¹³ C solid-state nuclear magnetic resonance (SSNMR) spectra shown in FIG. 7A. The method according to any one of claims 1 to 7,
Form A polymorph, characterized by ¹⁹ F SSNMR spectrum shown in FIG. 7B. (S) -2-((2-((S) -4- (difluoromethyl) -2) showing an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 10.8, 16.8, and 20.4 ° 2-theta Crystalline anhydride polymorph of oxooxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide Chain form D polymorph. The method of claim 9,
Form D polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 7.5, 8.6, 10.8, 16.8, 19.2, and 20.4 ° 2-theta. (S) -2-((2-((S) -4- (difluoromethyl) -2-oxo) showing an X-ray powder diffraction pattern with characteristic peaks at about 5.4, 10.5 and 25.2 ° 2-theta Crystalline trihydrate polymorph of oxazolidin-3-yl) -5,6-dihydrobenzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide Form B polymorph. A pharmaceutical composition comprising a therapeutically effective amount of the crystalline anhydride polymorph according to any one of claims 1 to 8 and a pharmaceutically acceptable carrier, lubricant, diluent or excipient. A pharmaceutical composition comprising a therapeutically effective amount of the crystalline anhydride polymorph according to claim 9 or 10 and a pharmaceutically acceptable carrier, glidant, diluent or excipient. A pharmaceutical composition comprising a therapeutically effective amount of the crystalline trihydrate polymorph according to claim 11 and a pharmaceutically acceptable carrier, lubricant, diluent or excipient. The method according to any one of claims 12 to 14,
Pharmaceutical compositions in the form of tablets. The method according to any one of claims 12 to 15,
A pharmaceutical composition, wherein the therapeutically effective amount is from 1 to 100 mg. The method according to any one of claims 12 to 16,
Pharmaceutical composition wherein the crystalline anhydride polymorph or crystalline trihydrate polymorph is milled. (S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-dihydrobenzo [] in ethanol or n-propanol [ f] After heating the slurry of imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide, the mixture is cooled to produce a characteristic peak at about 5.7 ° 2-theta. A method for producing a crystalline polymorph comprising forming a Form A crystalline polymorph exhibiting an X-ray powder diffraction pattern. The method of claim 18,
And Form A crystalline polymorph exhibiting an X-ray powder diffraction pattern with characteristic peaks at about 5.7, 11.4, 17.2, 19.0, 19.7, and 24.4 ° 2-theta. The method of claim 18 or 19,
Using ethanol or n-propanol with water. The method of claim 18 or 19,
Process of using ethanol or n-propanol without water. The method of claim 18 or 19,
(S) -2-((2-((S) -4- (difluoromethyl) -2-oxooxazolidin-3-yl) -5,6-di in ethanol in the presence of less than 40% water Heating a slurry of hydrobenzo [f] imidazo [1,2-d] [1,4] oxazinpin-9-yl) amino) propanamide. A method comprising administering to a subject in need thereof an effective amount of a crystalline polymorph according to any one of claims 1 to 11 or a pharmaceutical composition according to any one of claims 12 to 17. To cure cancer in a subject in need of treatment. The method according to any one of claims 1 to 17,
Crystalline polymorph or pharmaceutical composition for use in the treatment of cancer. Use of a crystalline polymorph according to any one of claims 1 to 11 or a pharmaceutical composition according to any one of claims 12 to 17 in the manufacture of a medicament for the treatment of cancer. Invention as described above.

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